Reproducibility and Other Issues in CSF proteomics to identify early biomarkers for neurodegenerative disease

We are performing CSF proteomics to identify early biomarkers for neurodegenerative disease using LC–MS/MS. Despite following standardized protocols, we are facing reproducibility and biological interpretation issues.

  1. Poor overlap of differentially expressed proteins between discovery and validation cohorts, even when using targeted MS.
  2. Ambiguous biological relevance, where statistically significant proteins lack clear mechanistic links to CNS pathology

Would alternative approaches (e.g., DIA-MS, extracellular vesicle enrichment, PTM-focused proteomics, or single-molecule methods) provide more reproducible insights?

What minimum standards should be enforced for CSF handling to improve cross-study comparability?

Thanks in advance

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